Methods to protect transplanted tissue from rejection

Abstract

The present invention includes compositions and methods for an HLA-A2 specific chimeric antigen receptor (CAR). In certain embodiments the HLA-A2 specific CAR is expressed on a T regulatory cell. In certain embodiments, the HLA-A2 specific CAR protects transplanted tissue from rejection.

What is claimed is: 
     
       1. A modified immune cell or precursor cell thereof, comprising a chimeric antigen receptor (CAR) having affinity for HLA-A2, wherein the CAR comprises, from N-terminus to C-terminus, a CD8 signal peptide, a HLA-A2 scFv binding domain comprising the amino acid sequence set forth in SEQ ID NO: 1, a CD8 hinge domain, a CD28 transmembrane domain, a CD28 intracellular domain, and a CD3 zeta intracellular domain, and wherein the HLA-A2 scFv binding domain cross-reacts with HLA-A28. 
     
     
       2. The modified immune cell or precursor cell thereof of  claim 1 , wherein the modified cell is a modified regulatory T cell. 
     
     
       3. The modified immune cell or precursor cell thereof of  claim 1 , wherein the modified cell is an autologous cell derived from a human. 
     
     
       4. An isolated nucleic acid comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR) having affinity for HLA-A2, wherein the CAR comprises, from N-terminus to C-terminus, a CD8 signal peptide, an HLA-A2 binding domain comprising the amino acid sequence set forth in SEQ ID NO: 1, a CD8 hinge domain, a CD28 transmembrane domain, a CD28 intracellular domain, and a CD3 zeta intracellular domain. 
     
     
       5. The isolated nucleic acid of  claim 4 , wherein the nucleic acid sequence comprises the sequence set forth in SEQ ID NO: 24. 
     
     
       6. An expression construct comprising the isolated nucleic acid of  claim 4 . 
     
     
       7. The expression construct of  claim 6 , wherein the expression construct comprises an EF-1α promoter, a rev response element (RRE), a woodchuck hepatitis virus posttranscriptional regulatory element (WPRE), and a cPPT sequence, and wherein the expression construct is a viral vector selected from the group consisting of a retroviral vector, a lentiviral vector, an adenoviral vector, and an adeno-associated viral vector. 
     
     
       8. The modified immune cell or precursor cell thereof of  claim 1 , wherein the CD8 signal peptide comprises the amino acid sequence set forth in SEQ ID NO: 13. 
     
     
       9. The modified immune cell or precursor cell thereof of  claim 1 , wherein the CD8 hinge domain comprises the amino acid sequence set forth in SEQ ID NO: 15. 
     
     
       10. The modified immune cell or precursor cell thereof of  claim 1 , wherein the CD28 transmembrane domain comprises the amino acid sequence set forth in SEQ ID NO: 17. 
     
     
       11. The modified immune cell or precursor cell thereof of  claim 1 , wherein the CD28 intracellular domain comprises the amino acid sequence set forth in SEQ ID NO: 19. 
     
     
       12. The modified immune cell or precursor cell thereof of  claim 1 , wherein the CD3 zeta intracellular domain comprises the amino acid sequence set forth in SEQ ID NO: 21. 
     
     
       13. The modified immune cell or precursor cell thereof of  claim 1 , wherein the CAR comprises the amino acid sequence set forth in SEQ ID NO: 23.